you can pre-incubate the primary antibody against Cyp17 with the same peptide used to obtain the antibody itself, see whether they sell it , it is called "blocking peptide". The antibody pre-adsorbed with the peptide should not recognize your protein anymore so the only band tha you see to disappear corresponds to your protein.
Thank you very much. I will contact them to know because I can't find the blocking peptide on their online catalog.
Is that the only way ? because if they don't sell it I don't really know what to do ?
another way would be use the same antibody on a Cyp17 Knock out mouse, at least a conditional knock out in the tissue of interest, there you should not see the band anymore and get the answer you need but I don't know whether there is such a transgenic mouse and whether it is vital...
I thought that I had to "prove" it for a publication but if this validation is not asked to publish indeed it's not vital. Because it would be a lot of work finding such a knock out mouse and negociation with their owners and so on...
Thank you for your answer it has been very helpfull.
I suppose you could just cut your band, strip the antibody, treat with trypsin and analyse peptides by mass spectrometry.
Attached you will find an article dealing with this, but sure you can obtain satisfactory results with other methods.
If you know what modification (glycosylation?) occurs, you could try a deglycosylation protocol (I used the EDEGLY kit from Sigma and it worked quite well) and run the treated and the control sample in parallel... the two bands should show at different moleculare weights.
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